It is well known that the expression results of any expression system are affected by many factors, including baculovirus expression system. As an expert in the field of protein expression, Creative BioMart has been pursuing the construction of excellent baculovirus expression system, and our research on system optimization has never stopped. By changing some parameters, the expression level of specific proteins can be optimized. We usually optimize the expression by changing five parameters.
Different expression approaches correspond to different vectors. Creative BioMart's vector gallery has included a wide range of vectors. Usually, in the initial stage of receiving expression services, our technicians will communicate with customers and select several suitable vectors or expression approaches for customers. Compare several vectors or expression methods to see which method can improves target yield and / or quality.
Although the polyhedrin and p10 promoters are inherently the strongest we can use, some more difficult to express proteins give better yields using an earlier promoter that are active when the cell is less compromised from virus infection. We can offer to compare the very late polyhedrin with the late p6.9. This decision needs to be made at the start of the project as it will affect which transfer vectors are used.
Host cell line will affect proteins yield, and quality can vary considerably. Creative BioMart offers several commonly used cell lines for our customers (click here to access to our cell line information). Of course, if customers want to use their own cell lines, we are willing to offer optimize service. In the stage of recombinant virus infecting insect cells, customers can choose more than one host cell for infection and rapid expression. Finally, the most suitable cell line was determined according to the expression effect.
Protein production and quality also vary with the MOI of the virus. Sometimes lower MOI can produce higher quality proteins. Therefore, we usually infect insect cells with different MOI viruses to determine the MOI value corresponding to the highest protein production (we normally compare MOIs of 0.5, 2 and 5).
Small shake cultures are set up in the appropriate cell lines. The BEVS recombinant gene product may or may not be secret. The peak expression of secretory protein usually appeared 30-72 hours after infection while that of non-secretory proteins usually reached the peak at 48-96 hours post-infection. To be on the safe side, after 24 hours post-infection, samples will be taken every 8-12 hours to determine the protein expression profile and the optimal harvest time.
Creative BioMart promises to update the progress in time, communicate with customers and exchange opinions in time. We are striving to deliver satisfactory results, please contact us for further assistance.